Effects of prophage regions in a plasmid carrying a carbapenemase gene on survival against antibiotic stress.

International Journal of Antimicrobial Agents
So Yeon Kim, Kwan Soo Ko

Abstract

We investigated the evolutionary importance of cryptic prophage elements in a blaNDM-1-bearing plasmid by exploring the effect of prophage regions on survival against antibiotic stress. While analysing a plasmid harboring an NDM-1-encoding gene in Klebsiella pneumoniae from South Korea, we found a prophage region within the plasmid. We constructed single-prophage knockout (KO) mutants by gene replacement. The intact plasmid and plasmids with deleted prophages were conjugated into Escherichia coli DH5α. Growth rate and antibiotic susceptibility were determined, and survival rates of strains were evaluated in the presence of antibiotics, such as imipenem, amikacin, gentamicin, cefotaxime, and piperacillin/tazobactam. A transcriptional response of sigma factor-coding genes (rpoS and rpoE) and reactive oxygen species (ROS)-related genes from different operons (soxS, fumC, oxyR, and katE) to a sub-inhibitory concentration of aminoglycosides was monitored by quantitative real-time polymerase chain reaction (qRT-PCR). The prophage region consists of four cryptic prophages of 16,795 bp and 19 coding DNA sequences. An Escherichia coli transconjugant carrying the plasmid with intact prophages showed increased survival during treatment wi...Continue Reading

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